Biology subjects

Naugler, D.

Publications and source records attributed to Naugler, D..

2 recordsLinked to original sources

bioRxiv2017

Notes On Synthesis Of Perdeutero-5-13C,5,5,5-Trifluoroisoleucine VI

The 13CF3 group is a promising label for heteronuclear (19F,13C) NMR studies of proteins. Desirable locations for this NMR spin label include the branched chain amino acid methyl groups. It is known that replacement of CH3 by CF3 at such locations preserves protein structure and function and enhances stability. 13CF3 may be introduced at the a position of isoleucine and incorporated biosynthetically in highly deuterated proteins. This paper reports our work in synthesis and purification of 5,5,5-trifluoroisoleucine, its perdeutero and 5-13C versions and of 2-13C-trifluoroacetate and its utility as a precursor for introduction of the 13CF3 group into proteins.\n\n\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=144 SRC=\"FIGDIR/small/140681_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (36K):\norg.highwire.dtl.DTLVardef@1531c5eorg.highwire.dtl.DTLVardef@16b8e29org.highwire.dtl.DTLVardef@423d0aorg.highwire.dtl.DTLVardef@b5951a_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

bioRxiv2017

NMR Analysis Of The Interaction Of Ethanol With The Nicotinic Acetylcholine Receptor

Ethanol exerts its actions in the central and peripheral nervous systems through the direct interactions with several proteins, including ligand-gated ion channels such as the nicotinic acetylcholine receptor (nAChR). The binding interaction between ethanol and sodium cholate solubilized nicotinic acetylcholine receptor protein can be detected through either NMR line broadening or T1 titration. In this paper, we examine the use of weighted Navon T1p analysis of T1 titration data for the estimation of the dissociation constant of ethanol for the nAChR. We show that Navon T1p analysis underestimates binding affinity. The application of rigorous limits for confidence intervals within a nonlinear regression analysis of this data provides a best estimate of Kd = 55 M at 4 {degrees}C. within an unsymmetrical 90% confidence interval of [0.5, 440 M]. Accordingly, the best estimate of the binding free energy is {Delta}G0, = -5.4 Kcal/mole within a 90% confidence interval of [-8.0, -4.3 Kcal/mole],relative to conventional standard states.

molecular biology↗